AgNO3 DNA Staining Procedure Questions (David Atherton, are you there?)
Mr. D.J. Atherton
datherto at crc.ac.uk
Fri Jun 18 05:25:02 EST 1993
Hi Tim, Netters
My deepest and most humble apologies if I wrote to develop in Na2CO3/formamide
Developing solution should read:-
Develop in 2% Na2CO3/0.2% **FORMALDEHYDE**
I really must learn to lay off the 100% EtOH :-).
Can't think why your mail bounced except perhaps the @address I put is in
JANET format, ie the other way around from Internet. Failing that try just
'reply'ing when reading the mail and go ad lib.
datherton at uk.ac.ox.path.vax
PS. Had this sent to me, thought you would like it as well.
(Sorry about cutting the .sig Ed!)
And another, from personal comm:
RNA / DNA SILVER STAIN PROTOCOL FOR ACRYLAMIDE GELS
FROM: Moshe Balas, Volcani Centre, Rehovot, Israel
BY: Ed Rybicki, Dept Microbiology, University of Cape Town, Aug 1992
WORKS VERY WELL FOR VIROIDS, dsRNAs, AND REPUTEDLY FOR DNA ALSO
1. Soak in 30% EtOH / 10% acetic acid 10 min on bench, can
leave up to O/N.
2. Soak 10% EtOH 2x10 min, wash in ddH2O 3x5 min.
3. Soak in 6 mM AgNO3 for 30 min (50 ml/gel), then quick rinse
4. Rinse rapidly in fresh 2.5% Na2CO3 / 0.02% HCHO, then fresh
soln. for 2 min or longer: develop in same solution till
bands appear dense enough.
5. Stop with 1% acetic and keep in same solution.
NB: CAN DESTAIN GELS WITH 2% AMMONIUM PEROXYDISULPHATE
NNB: CAN SHRINK GELS FOR STORAGE BY SOAKING IN ABSOLUTE EtOH,
THEN ACETONE, THEN AIR-DRYING
1. Soak polyacrylamide AFTER ELECTROPHORESIS in 50% EtOH / 10%
acetic acid 1-3 hr or or overnight.
2. Soak in 10% EtOH / 1% acetic acid 1 hr, rinse 3x in ddH2O.
3. Soak in 12 mM AgNO3 1 hr, rinse 3x5 min in ddH2O.
4. Rinse rapidly in fresh 0.75 M KOH / 0.28% HCHO, leave to
develop in more of same solution till bands appear at
sufficient contrast / density.
5. Add XS H2O to stop or stop with 0.07 M Na2CO3 or 5% acetic
acid. Store in 1% acetic.
| Ed Rybicki, PhD |
| (ed at micro.uct.ac.za) |
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